Profiling anti-influenza activities of the pathogen recognition receptor RIG-I: Novel targets for antiviral intervention strategies

分析病原体识别受体 RIG-I 的抗流感活性：抗病毒干预策略的新靶点

• 批准号: 319869332
• 负责人: Professor Dr. Stefan Bauer
• 金额: --
• 依托单位: Institut für Virologie
• 依托单位国家: 德国
• 项目类别: Clinical Research Units
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/319869332?language=en
• 关键词: Profiling; anti; influenza; activities; pathogen

RIG-I is a cellular virus sensor that binds the 5’ppp-dsRNA panhandle of Influenza A virus (FLUAV). RIG-I then switches conformation and can exert a direct antiviral effect by clamping onto the panhandle and disassembling the polymerase complex, but normally it also triggers antiviral signaling leading to type I interferon (IFN) production. We had generated mutants of RIG-I that reduce viral RNA synthesis despite being deficient in antiviral signaling. The antiviral mechanism of these RIG-I mutants will be explored and optimized by defining the minimally required domain and by further modifications. We already have created mice bearing one such RIG-I mutant (K271A), and will compare their FLUAV resistance with wt and RIG-I KO mice. Paliperidone, an FDA-approved antipsychotic predicted by Patel & Kukol (Virology 2017) to bind PB2, was shown to mimic the direct antiviral activity of RIG-I as it diminished the binding of PB2 to nucleocapsids and reduced RNA synthesis of the PR8/34 strain. We will test Paliperidone against a variety of FLUAV strains in cell lines and primary human airway cells, investigate a potential involvement of RIG-I in its antiviral effect, and evaluate its interplay with RIG-I agonists. Also, we identified rRNA-derived self-RNA fragments as new RIG-I ligands. These RIG-I stimulators can be generated by the OAS/ RNase L that in turn can be activated by dsRNA and the glycolysis intermediate Fructose-1,6-bisphosphate (F16bP). This possible link between an endogenous intermediate of glycolysis and innate immunity will be followed up in OAS/RNase L knockout cells. Moreover, the influence of the metabolic state of macrophages on RNase L activation, self-RNA fragment generation and subsequent RIG-I-mediated antiviral activity will be investigated.

RIG-I 是一种细胞病毒传感器，可结合甲型流感病毒 (FLUAV) 的 5'ppp-dsRNA 狭长区域。然后，RIG-I 会改变构象，并可以通过夹住狭长区域并分解聚合酶复合物来发挥直接抗病毒作用，但通常它也会触发抗病毒信号传导，导致 I 型干扰素 (IFN) 的产生。我们生成了 RIG-I 突变体，尽管其抗病毒信号传导存在缺陷，但仍能减少病毒 RNA 合成。这些 RIG-I 突变体的抗病毒机制将通过定义所需的最低结构域和进一步修改来探索和优化。我们已经创建了携带这种 RIG-I 突变体 (K271A) 的小鼠，并将其与 wt 和 RIG-I KO 小鼠进行 FLUAV 抗性比较。帕利哌酮是 FDA 批准的抗精神病药物，Patel & Kukol (Virology 2017) 预测它可以与 PB2 结合，它被证明可以模仿 RIG-I 的直接抗病毒活性，因为它减少了 PB2 与核衣壳的结合，并减少了 PR8/34 病毒株的 RNA 合成。我们将在细胞系和原代人气道细胞中针对多种 FLUAV 毒株测试帕潘立酮，研究 RIG-I 对其抗病毒作用的潜在参与，并评估其与 RIG-I 激动剂的相互作用。此外，我们还鉴定了 rRNA 衍生的自身 RNA 片段作为新的 RIG-I 配体。这些 RIG-I 刺激剂可由 OAS/RNase L 产生，而 OAS/RNase L 又可由 dsRNA 和糖酵解中间体果糖-1,6-二磷酸 (F16bP) 激活。糖酵解内源性中间体与先天免疫之间可能存在的联系将在 OAS/RNase L 敲除细胞中进行追踪。此外，还将研究巨噬细胞的代谢状态对 RNase L 激活、自身 RNA 片段生成以及随后的 RIG-I 介导的抗病毒活性的影响。